SNP typing method and kit

A typing method and kit technology, applied in the field of DNA detection, can solve problems such as false signals, and achieve the effect of reducing false signals

Inactive Publication Date: 2017-06-13
GUANGZHOU KANGXINRUI GENE HEALTH TECH CO LTD
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Problems solved by technology

[0006] The purpose of the present invention is to provide a SNP typing method and kit with higher accuracy for the detection of SNP sites in CYP2C9, VKORC1 and CYP2C19 genes, aiming to solve the high proportion of errors in the sequencing results in the prior art Signal technical issues

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no. 3 example

[0083] Based on the third embodiment, the present invention also proposes a fourth embodiment, the ligation reaction in the step B1 is carried out in a cutting-ligation reaction system, and the cutting-ligation reaction system includes: ligase, cleavage agent, immobilized First linker and ligation buffer on microspheres;

[0084] The cleaving agent is used to specifically cut the cleavable site or excisable sequence in the nucleic acid fragment to form the first cohesive end;

[0085] The first linker is a nucleic acid molecule containing a second cohesive end that is completely complementary to the first cohesive end.

[0086] It should be noted that the cleavage-ligation reaction system is a reaction system in which the ligase and the cleavage agent are active at the same time.

[0087] Through the above-mentioned special design of PCR amplification primers and cutting-ligation reaction system, in the process of library construction, the cutting and ligation reactions after...

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Abstract

The invention relates to the technical field of DNA detection and provides an SNP typing method and kit. A to-be-tested SNP site comprises SNP sites in CYP2C9, VKORC1 and CYP2C19 genes. The SNP typing method comprises the following steps: replacing a fluorescently-labeled oligonucleotide probe with a non-fluorescently-labeled oligonucleotide probe, performing one-time connection sequencing reaction on to-be-sequenced library molecules and then obtaining sequence information of the to-be-tested SNP site through connection sequencing. The SNP typing kit comprises a CYP2C9 specific primer pair, a VKORC1 specific primer pair, a CYP2C19 specific primer pair and the non-fluorescently-labeled oligonucleotide probe. The SNP typing method and kit provided by the invention have the benefit that through sealing non-specific binding sites in the to-be-sequenced library molecules, a false signal in a result obtained by the connection sequencing is reduced, so that the accuracy of SNP site detection is improved.

Description

technical field [0001] The invention relates to the technical field of DNA detection, in particular to a SNP typing method and a kit. Background technique [0002] Single nucleotide polymorphism (Single Nucleotide Polymorphism, SNP) refers to the existence of conversion, transversion, insertion, deletion and other changes at a specific nucleotide position in DNA in the genome. At present, common SNP typing detection technologies at home and abroad include the following, high resolution melting curve detection (high resolution melting, HRM), SNP typing method based on chip technology, allele-specific amplification method (Amplification Refractory Mutation System Real Time PCR, ARMS-RT PCR), Taqman fluorescent probe method, mass spectrometry, high performance liquid chromatography and gene sequencing, etc. [0003] Among them, the gene sequencing method also includes the first-generation sequencing technology based on Sanger technology, and the second-generation high-throughp...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q2563/149C12Q2525/191C12Q2565/518
Inventor 盛司潼邓波
Owner GUANGZHOU KANGXINRUI GENE HEALTH TECH CO LTD
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